CN103193880A - Preparation method of HCG (Human Chorionic Gonadotropin) crude product - Google Patents
Preparation method of HCG (Human Chorionic Gonadotropin) crude product Download PDFInfo
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- CN103193880A CN103193880A CN201310120891XA CN201310120891A CN103193880A CN 103193880 A CN103193880 A CN 103193880A CN 201310120891X A CN201310120891X A CN 201310120891XA CN 201310120891 A CN201310120891 A CN 201310120891A CN 103193880 A CN103193880 A CN 103193880A
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- 239000012043 crude product Substances 0.000 title claims abstract description 48
- 238000002360 preparation method Methods 0.000 title claims abstract description 28
- 102000011022 Chorionic Gonadotropin Human genes 0.000 title abstract description 8
- 108010062540 Chorionic Gonadotropin Proteins 0.000 title abstract description 8
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims abstract description 82
- 238000000108 ultra-filtration Methods 0.000 claims abstract description 59
- 210000002700 urine Anatomy 0.000 claims abstract description 44
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims abstract description 13
- 238000003756 stirring Methods 0.000 claims abstract description 13
- 238000005406 washing Methods 0.000 claims abstract description 10
- 208000030507 AIDS Diseases 0.000 claims description 15
- 241000700721 Hepatitis B virus Species 0.000 claims description 15
- 241000700605 Viruses Species 0.000 claims description 15
- 239000000843 powder Substances 0.000 claims description 12
- 239000001110 calcium chloride Substances 0.000 claims description 11
- 229910001628 calcium chloride Inorganic materials 0.000 claims description 11
- 238000002156 mixing Methods 0.000 claims description 11
- 238000001556 precipitation Methods 0.000 claims description 11
- 238000001291 vacuum drying Methods 0.000 claims description 11
- 239000000047 product Substances 0.000 claims description 10
- 239000007788 liquid Substances 0.000 claims description 9
- 239000012528 membrane Substances 0.000 claims description 9
- 238000001471 micro-filtration Methods 0.000 claims description 9
- 238000002203 pretreatment Methods 0.000 claims description 9
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 7
- 238000005057 refrigeration Methods 0.000 claims description 6
- 238000001223 reverse osmosis Methods 0.000 claims description 6
- 230000002000 scavenging effect Effects 0.000 claims description 6
- 238000005304 joining Methods 0.000 claims description 3
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 abstract description 12
- WXMKPNITSTVMEF-UHFFFAOYSA-M sodium benzoate Chemical compound [Na+].[O-]C(=O)C1=CC=CC=C1 WXMKPNITSTVMEF-UHFFFAOYSA-M 0.000 abstract description 8
- 239000004299 sodium benzoate Substances 0.000 abstract description 8
- 235000010234 sodium benzoate Nutrition 0.000 abstract description 8
- 229940084986 human chorionic gonadotropin Drugs 0.000 abstract description 6
- 239000006228 supernatant Substances 0.000 abstract description 4
- 238000006243 chemical reaction Methods 0.000 abstract description 3
- 239000003814 drug Substances 0.000 abstract description 2
- 238000001035 drying Methods 0.000 abstract 2
- 229940079593 drug Drugs 0.000 abstract 1
- 230000001376 precipitating effect Effects 0.000 abstract 1
- 239000000243 solution Substances 0.000 description 55
- 238000000034 method Methods 0.000 description 16
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 13
- 210000004379 membrane Anatomy 0.000 description 7
- 238000005516 engineering process Methods 0.000 description 5
- 238000012360 testing method Methods 0.000 description 5
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 4
- 229960004407 chorionic gonadotrophin Drugs 0.000 description 4
- 230000035935 pregnancy Effects 0.000 description 4
- WPYMKLBDIGXBTP-UHFFFAOYSA-N benzoic acid Chemical compound OC(=O)C1=CC=CC=C1 WPYMKLBDIGXBTP-UHFFFAOYSA-N 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 230000000052 comparative effect Effects 0.000 description 3
- 238000000605 extraction Methods 0.000 description 3
- 230000006870 function Effects 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 239000002594 sorbent Substances 0.000 description 3
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 2
- 102000003886 Glycoproteins Human genes 0.000 description 2
- 108090000288 Glycoproteins Proteins 0.000 description 2
- RJKFOVLPORLFTN-LEKSSAKUSA-N Progesterone Chemical compound C1CC2=CC(=O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H](C(=O)C)[C@@]1(C)CC2 RJKFOVLPORLFTN-LEKSSAKUSA-N 0.000 description 2
- 230000004720 fertilization Effects 0.000 description 2
- 210000002826 placenta Anatomy 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 230000028327 secretion Effects 0.000 description 2
- SXHWYMWRVLSNAW-UHFFFAOYSA-L C(C)O.[Cl-].[Ca+2].C(C)(=O)[O-].[Na+] Chemical compound C(C)O.[Cl-].[Ca+2].C(C)(=O)[O-].[Na+] SXHWYMWRVLSNAW-UHFFFAOYSA-L 0.000 description 1
- 208000006332 Choriocarcinoma Diseases 0.000 description 1
- 206010011498 Cryptorchism Diseases 0.000 description 1
- 208000008899 Habitual abortion Diseases 0.000 description 1
- 206010062767 Hypophysitis Diseases 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- 206010046788 Uterine haemorrhage Diseases 0.000 description 1
- DPDMMXDBJGCCQC-UHFFFAOYSA-N [Na].[Cl] Chemical compound [Na].[Cl] DPDMMXDBJGCCQC-UHFFFAOYSA-N 0.000 description 1
- 238000010923 batch production Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 210000004252 chorionic villi Anatomy 0.000 description 1
- 210000004246 corpus luteum Anatomy 0.000 description 1
- 201000000160 cryptorchidism Diseases 0.000 description 1
- 238000013016 damping Methods 0.000 description 1
- 210000003785 decidua Anatomy 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 230000018044 dehydration Effects 0.000 description 1
- 238000006297 dehydration reaction Methods 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 230000001076 estrogenic effect Effects 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 231100000535 infertility Toxicity 0.000 description 1
- 208000000509 infertility Diseases 0.000 description 1
- 230000036512 infertility Effects 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 231100000545 luteal phase defect Toxicity 0.000 description 1
- 230000035800 maturation Effects 0.000 description 1
- 238000010999 medical injection Methods 0.000 description 1
- 239000003960 organic solvent Substances 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 238000009597 pregnancy test Methods 0.000 description 1
- 229960003387 progesterone Drugs 0.000 description 1
- 239000000186 progesterone Substances 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000004062 sedimentation Methods 0.000 description 1
- 230000036299 sexual function Effects 0.000 description 1
- 239000001632 sodium acetate Substances 0.000 description 1
- 229960004249 sodium acetate Drugs 0.000 description 1
- 235000017281 sodium acetate Nutrition 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
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- Peptides Or Proteins (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The invention belongs to the technical field of biological medicines and in particular relates to a preparation method of an HCG (Human Chorionic Gonadotropin) crude product. The preparation method of the HCG crude product comprises the following steps of: a, collecting urine of a pregnant woman and obtaining a concentrated solution through ultra-filtration; b, adding anhydrous calcium chloride with concentration to 0.5%-0.8% to the concentrated solution, and dissolving the anhydrous calcium chloride by stirring; c, adding the concentrated solution in the step b to acetone, which is 2-3 times the volume of the concentrated solution, for stewing for 10-16 hours, wherein the temperature of the acetone is 4 DEG C and the acetone added with the concentrated solution is stored under 4 DEG C; and d, siphoning the supernatant in the step c and washing and precipitating the supernatant twice by acetone, and drying or drying in vacuum to obtain the HCG crude product. Compared with the conventional preparation method of the sodium benzoate, the HCG crude product prepared by the preparation method disclosed by the invention does not need sodium benzoate and hydrochloric acid, so that the cost is reduced. In the preparation method disclosed by the invention, the HCG loss caused by an intermediate reaction link is reduced.
Description
Technical field
The invention belongs to technology biological medicine technology field, be specifically related to a kind of preparation method of HCG crude product.
Background technology
HCG is human chorionic gonadotrophin, is the lead-in letter abbreviations of three English of English human chorionic gonadotrophin, is that it is made up of α and the dimeric glycoprotein of β by a kind of glycoprotein of trophocyte's secretion of placenta.
HCG just enters female blood and fast breeding before and after the 6th day until the 8th week in pregnancy period is slowly reduced concentration then up to the 18th～20 week at after fertilization, keeps then about 10 days, and begins descend (but still being higher than normal level).Complete HCG all is the plamoditrophoblast generation by the placental villi film.Its major function stimulates corpus luteum exactly, is conducive to oestrogenic hormon and Progesterone continuous release, to promote the formation of uterine decidua, makes the placenta growth maturation.Modern times think that HCG is produced by trophoderm transitional-cell and syncytium.HCG began secretion in about 6 days at after fertilization, peaked in 60-70 days.Preceding 8 weeks increment in gestation is very fast, to keep gestation.After about pregnant 8 weeks, HCG descends gradually, reaches relatively stable up to about 20 weeks.
The HCG proterties powder that is white in color, soluble in water, be insoluble to organic solvents such as ethanol, acetone, ether, pH3.2-3.3, dry product is more stable, be mainly used in the low excessively cryptorchidism of sexual function due to male sex's hypophysis function deficiency clinically, be used for the sick and habitual abortion of the function row uterine hemorrhage that caused by luteal phase defect and HMG contract and can bring out and ovulate and treat infertility.Another important use of HCG is preparation Clearblue Pregnancy Test box radioimmunity reagent, and in order to the diagnosis of gestation check and choriocarcinoma, HCG is medically having important purposes.
The traditional preparation process method of HCG is: collect pregnant woman urine, in urine, add Sodium Benzoate, add hydrochloric acid again, fully reaction generates phenylformic acid, HCG is adsorbed, add edible ethanol again HCG is precipitated, dry HCG again, the shortcoming of this method is, the step of this method is various, and the HCG loss in the urine is bigger; And having used Sodium Benzoate and hydrochloric acid in this method, its cost is higher, and the multiple new process need consumption of calorie of hydrochloric acid.
In the CN1302818A disclosed method, adopt high salt extracting and the less salt back suction technology of sodium-acetate to obtain the HCG intermediate, separating and purifying technology through the SP-dextrane gel makes a kind of highly purified human chorionic gonadotrophin, makes medical injection HCG elaboration raw material through aseptic pyrogen-free processing.Its concrete technical process is: intermediate makes workshop section and pure product HCG makes workshop section, intermediate is made in workshop section's process, take by weighing the HCG crude product, put in the damping fluid of sodium-acetate calcium chloride ethanol of sodium-chlor, stir and extract, standing over night siphon supernatant liquor, residue continue to extract the centrifuging and taking supernatant liquor; Adding ethanol continues to extract; Adopt pre-cold acetone dehydration final vacuum drying, get intermediate; Cross chromatographic column to the HCG purification of intermediate, collect, add ethanol sedimentation, tire and be the pure product of HCG of 3000-3500iu/mg.The shortcoming of this method is: the reagent of employing is more, and step is comparatively loaded down with trivial details, and has adopted organic reagent ethanol in the method, has increased the step of solvent recuperation.
CN1047507A has disclosed a kind of sorbent material and method of extraction of human chorionic gonadotropin, the concrete steps of this method are: use diatomite as sorbent material, from early, mid-term (2-6 month) pregnant woman's freshly voided urine, at extraction of human chorionic gonadotropin below 37 ℃.Be specially: regulate pregnant woman urine pH value, add diatomite adsorption, get diatomite dress post, use the elutriant wash-out, add alcohol precipitation, make human chorionic gonadotrophin through dehydrating.The shortcoming of this method is: adopt diatomite as sorbent material, can adsorb a part of HCG, bring the loss of HCG.
Summary of the invention
In order to solve above-mentioned technical problem, the method for the preparation of the HCG crude product that the invention provides that a kind of cost is low, step is simple, is easy to suitability for industrialized production.
The preparation method of HCG crude product comprises following step:
A. collect pregnant woman urine, adopt 40 mesh filter screens to filter; Adopt the microfiltration equipment pre-treatment to filter, ultrafiltration obtains concentrated solution in ultrafiltration apparatus again, and the condition of ultrafiltration is: pH3.8-4.0, the molecular weight cut-off of ultra-filtration membrane are 10000Da, till ultrafiltration to concentrated solution is the 4-5% of former urine weight;
B. in concentrated solution, add and account for concentrated solution and heavily be the Calcium Chloride Powder Anhydrous of 0.5-0.8%, stirring and dissolving;
C. again the concentrated solution among the step b is joined being pre-chilled in 4 ℃ the acetone of 2-3 times of volume and left standstill 6-15 hour, and place 4 ℃ to preserve down the mixing solutions of concentrated solution and acetone;
D. the upper strata liquid among the siphon step c, and with twice of washing with acetone precipitation; Drain or vacuum-drying gets the HCG crude product.
Among the above-mentioned step a, after the ultrafiltration urine, clean ultrafiltration apparatus with reverse osmosis water, next urine is incorporated in scavenging solution refrigeration into, the effective ingredient in the urine can be cleaned out more completely, reduces the loss in extracting.
Among the above-mentioned step b, adding war concentrated solution gross weight is 0.65% Calcium Chloride Powder Anhydrous in concentrated solution, stirring and dissolving.
Preferably, among the above-mentioned step c, be specially: left standstill 12 hours joining in the concentrated solution among the step b in the acetone of 2.5 times of volumes, described acetone is to be pre-chilled to 4 ℃ acetone again, and places 4 ℃ to preserve down the mixing solutions of concentrated solution and acetone.
Among the above-mentioned step a, in the pregnant woman urine of collection, must not detect virus of AIDS, hepatitis B virus HBs-Ab, HBc-Ab, Hbe-Ab.
Among the above-mentioned step a, must not detect virus of AIDS, hepatitis B virus HBs-Ab, HBc-Ab, Hbe-Ab in the resulting HCG crude product finished product.
Among the step a, in 4-10 ℃ interval range, be conducive to the extraction of HCG crude product.
The preparation method of HCG crude product comprises following step:
A. collect pregnant woman urine, adopt 40 mesh filter screens to filter; Adopt the microfiltration equipment pre-treatment to filter, ultrafiltration obtains concentrated solution in ultrafiltration apparatus again, and the condition of ultrafiltration is: pH3.9, the molecular weight cut-off of ultra-filtration membrane are 10000Da, ultrafiltration to concentrated solution be former urine weight 4.5% till;
B. add in concentrated solution that to account for concentrated solution heavily be 0.6% Calcium Chloride Powder Anhydrous, stirring and dissolving;
C. again the concentrated solution among the step b is joined being pre-chilled in 4 ℃ the acetone of 2.5 times of volumes and left standstill 8 hours, and place 4 ℃ to preserve down the mixing solutions of concentrated solution and acetone;
D. the upper strata liquid among the siphon step c, and with twice of washing with acetone precipitation; Drain or vacuum-drying gets the HCG crude product.
Beneficial effect of the present invention is, adopts above method to prepare the HCG crude product, compares with the preparation method of traditional employing Sodium Benzoate, need not to adopt Sodium Benzoate and hydrochloric acid, reduced cost; And in the method for the present invention, reduced because the loss of the HCG that the intermediate reaction link is brought.
Embodiment
Below in conjunction with specific embodiment the present invention is done further explanation, so that those skilled in the art more understands the present invention, but therefore do not limit the present invention.
Embodiment 1
The preparation method of HCG crude product comprises following step:
A. collect pregnant woman urine, in this pregnant woman urine, do not detect virus of AIDS, hepatitis B virus HBs-Ab, HBc-Ab, Hbe-Ab, adopt 40 mesh filter screens to filter; Adopt the microfiltration equipment pre-treatment to filter, ultrafiltration obtains concentrated solution in ultrafiltration apparatus again, and the condition of ultrafiltration is: pH3.9, the molecular weight cut-off of ultra-filtration membrane are 10000Da, and ultrafiltration to concentrated solution is about 4% of former urine weight;
Clean ultrafiltration apparatus with reverse osmosis water after the ultrafiltration, next urine is incorporated in scavenging solution refrigeration into;
B. in concentrated solution, add concentration and be 0.65% Calcium Chloride Powder Anhydrous, stirring and dissolving;
C. again the concentrated solution among the step b is joined being pre-chilled in 4 ℃ the acetone of 2.5 times of volumes and leave standstill more than 6 hours, and place 4 ℃ to preserve down the mixing solutions of concentrated solution and acetone;
D. the upper strata liquid among the siphon step c, and with twice of washing with acetone precipitation; Vacuum-drying gets the HCG crude product, after testing, does not detect virus of AIDS, hepatitis B virus HBs-Ab, HBc-Ab, Hbe-Ab in the HCG crude product.
Adopt the method among the above embodiment to prepare the HCG crude product, its experimental result is as follows:
Comparative Examples
Collect pregnant woman urine, in this pregnant woman urine, do not detect virus of AIDS, hepatitis B virus HBs-Ab, HBc-Ab, Hbe-Ab; Add Sodium Benzoate in urine, the add-on of Sodium Benzoate is 2.2%, adds hydrochloric acid to pH value 3.9 again and fully reacts, and generates phenylformic acid, and phenylformic acid adsorbs HCG, discards useless urine after 2 hours and dries precipitation.To dry thing again and add edible ethanol, vacuum-drying gets the HCG crude product to make HCG precipitate also, and the ratio of crude product is lived and is 5-8IU/mg.
Data from above form are come as can be seen, crude product among the embodiment 1 is than living greater than the crude product in the Comparative Examples than living, and the production cost of per 50 grams of the HCG product among the embodiment 1 is than production cost in the Comparative Examples low about 15%, every batch production time want the major general near half, this shows, the preparation method of HCG crude product of the present invention, its cost is low, time is few, the efficient height.
Embodiment 2
The preparation method of HCG crude product comprises following step:
A. collect pregnant woman urine, in this pregnant woman urine, do not detect virus of AIDS, hepatitis B virus HBs-Ab, HBc-Ab, Hbe-Ab, adopt 40 mesh filter screens to filter; Adopt the microfiltration equipment pre-treatment to filter, ultrafiltration obtains concentrated solution in ultrafiltration apparatus again, and the condition of ultrafiltration is: pH3.8, the molecular weight cut-off of ultra-filtration membrane are 10000Da, and ultrafiltration to concentrated solution is about 4.5% of former urine weight;
Clean ultrafiltration apparatus with reverse osmosis water after the ultrafiltration, next urine is incorporated in scavenging solution refrigeration into;
B. in concentrated solution, add concentration and be 0.6% Calcium Chloride Powder Anhydrous, stirring and dissolving;
C. again the concentrated solution among the step b is joined being pre-chilled in 4 ℃ the acetone of 3 times of volumes and leave standstill more than 10 hours, and place 4 ℃ to preserve down the mixing solutions of concentrated solution and acetone;
D. the upper strata liquid among the siphon step c, and with twice of washing with acetone precipitation; Vacuum-drying gets the HCG crude product, after testing, does not detect virus of AIDS, hepatitis B virus HBs-Ab, HBc-Ab, Hbe-Ab in the finished product HCG crude product.
Embodiment 3
The preparation method of HCG crude product comprises following step:
A. collect pregnant woman urine, in this pregnant woman urine, do not detect virus of AIDS, hepatitis B virus HBs-Ab, HBc-Ab, Hbe-Ab, adopt 40 mesh filter screens to filter; Adopt the microfiltration equipment pre-treatment to filter, ultrafiltration obtains concentrated solution in ultrafiltration apparatus again, and the condition of ultrafiltration is: pH4.0, the molecular weight cut-off of ultra-filtration membrane are 10000Da, and ultrafiltration to concentrated solution is about 4% of former urine weight;
Clean ultrafiltration apparatus with reverse osmosis water after the ultrafiltration, next urine is incorporated in scavenging solution refrigeration into;
B. in concentrated solution, add concentration and be 0.7% Calcium Chloride Powder Anhydrous, stirring and dissolving;
C. leave standstill more than 10 hours joining being pre-chilled in 4 ℃ the acetone of 3 times of volumes in the concentrated solution among the step b again, and place 4 ℃ to preserve down the mixing solutions of concentrated solution and acetone;
D. the upper strata liquid among the siphon step c, and with twice of washing with acetone precipitation; Vacuum-drying gets the HCG crude product, after testing, does not detect virus of AIDS, hepatitis B virus HBs-Ab, HBc-Ab, Hbe-Ab in the finished product HCG crude product.
Embodiment 4
The preparation method of HCG crude product comprises following step:
A. collect pregnant woman urine, in this pregnant woman urine, do not detect virus of AIDS, hepatitis B virus HBs-Ab, HBc-Ab, Hbe-Ab, adopt 40 mesh filter screens to filter; Adopt the microfiltration equipment pre-treatment to filter, ultrafiltration obtains concentrated solution in ultrafiltration apparatus again, and the condition of ultrafiltration is: pH3.8, the molecular weight cut-off of ultra-filtration membrane are 10000Da, and ultrafiltration to concentrated solution is about 5% of former urine weight;
Clean ultrafiltration apparatus with reverse osmosis water after the ultrafiltration, next urine is incorporated in scavenging solution refrigeration into;
B. in concentrated solution, add concentration and be 0.7% Calcium Chloride Powder Anhydrous, stirring and dissolving;
C. again the concentrated solution among the step b is joined being pre-chilled in 4 ℃ the acetone of 2.5 times of volumes and leave standstill more than 10 hours, and place 6 ℃ to preserve down the mixing solutions of concentrated solution and acetone;
D. the upper strata liquid among the siphon step c, and with twice of washing with acetone precipitation; Vacuum-drying gets the HCG crude product, after testing, does not detect virus of AIDS, hepatitis B virus HBs-Ab, HBc-Ab, Hbe-Ab in the finished product HCG crude product.
Embodiment 5
The preparation method of HCG crude product comprises following step:
A. collect pregnant woman urine, in this pregnant woman urine, do not detect virus of AIDS, hepatitis B virus HBs-Ab, HBc-Ab, Hbe-Ab, adopt 40 mesh filter screens to filter; Adopt the microfiltration equipment pre-treatment to filter, ultrafiltration obtains concentrated solution in ultrafiltration apparatus again, and the condition of ultrafiltration is: pH3.9, the molecular weight cut-off of ultra-filtration membrane are 10000Da, ultrafiltration to concentrated solution be former urine weight 4.5% till;
B. add in concentrated solution that to account for concentrated solution heavily be 0.6% Calcium Chloride Powder Anhydrous, stirring and dissolving;
C. again the concentrated solution among the step b is joined being pre-chilled in 4 ℃ the acetone of 2.5 times of volumes and left standstill 8 hours, and place 4 ℃ to preserve down the mixing solutions of concentrated solution and acetone;
D. the upper strata liquid among the siphon step c, and with twice of washing with acetone precipitation; Drain or vacuum-drying gets the HCG crude product, after testing, do not detect virus of AIDS, hepatitis B virus HBs-Ab, HBc-Ab, Hbe-Ab in the finished product HCG crude product.
Claims (8)
1.HCG the preparation method of crude product comprises following step:
A. collect pregnant woman urine, adopt 40 mesh filter screens to filter; Adopt the microfiltration equipment pre-treatment to filter, ultrafiltration obtains concentrated solution in ultrafiltration apparatus again, and the condition of ultrafiltration is: pH3.8-4.0, the molecular weight cut-off of ultra-filtration membrane are 10000Da, till ultrafiltration to concentrated solution is the 4-5% of former urine weight;
B. in concentrated solution, add and account for concentrated solution and heavily be the Calcium Chloride Powder Anhydrous of 0.5-0.8%, stirring and dissolving;
C. again the concentrated solution among the step b is joined being pre-chilled in 4 ℃ the acetone of 2-3 times of volume and left standstill 6-15 hour, and place 4 ℃ to preserve down the mixing solutions of concentrated solution and acetone;
D. the upper strata liquid among the siphon step c, and with twice of washing with acetone precipitation; Drain or vacuum-drying gets the HCG crude product.
2. the preparation method of HCG crude product as claimed in claim 1 is characterized in that, among the described step a, after the ultrafiltration urine, cleans ultrafiltration apparatus with reverse osmosis water, and next urine is incorporated in scavenging solution refrigeration into.
3. the preparation method of HCG crude product as claimed in claim 1 is characterized in that, among the described step b, adding concentration is 0.65% Calcium Chloride Powder Anhydrous in concentrated solution, stirring and dissolving.
4. the preparation method of HCG crude product as claimed in claim 1, it is characterized in that, be specially among the described step c: left standstill 12 hours joining in the concentrated solution among the step b in the acetone of 2.5 times of volumes again, described acetone is to be pre-chilled to 4 ℃ acetone, and places 4 ℃ to preserve down the mixing solutions of concentrated solution and acetone.
5. the preparation method of HCG crude product as claimed in claim 1 is characterized in that, among the described step a, in the described pregnant woman urine, must not detect virus of AIDS, hepatitis B virus HBs-Ab, HBc-Ab, Hbe-Ab.
6. the preparation method of HCG crude product as claimed in claim 1 is characterized in that, among the described step a, must not detect virus of AIDS, hepatitis B virus HBs-Ab, HBc-Ab, Hbe-Ab in the described finished product HCG crude product.
7. the preparation method of HCG crude product as claimed in claim 1 is characterized in that, among the described step a, the temperature of ultrafiltration is 4-10 ℃.
8. the preparation method of HCG crude product as claimed in claim 1 comprises following step:
A. collect pregnant woman urine, adopt 40 mesh filter screens to filter; Adopt the microfiltration equipment pre-treatment to filter, ultrafiltration obtains concentrated solution in ultrafiltration apparatus again, and the condition of ultrafiltration is: pH3.9, the molecular weight cut-off of ultra-filtration membrane are 10000Da, ultrafiltration to concentrated solution be former urine weight 4.5% till;
B. add in concentrated solution that to account for concentrated solution heavily be 0.6% Calcium Chloride Powder Anhydrous, stirring and dissolving;
C. again the concentrated solution among the step b is joined being pre-chilled in 4 ℃ the acetone of 2.5 times of volumes and left standstill 8 hours, and place 4 ℃ to preserve down the mixing solutions of concentrated solution and acetone;
D. the upper strata liquid among the siphon step c, and with twice of washing with acetone precipitation; Drain or vacuum-drying gets the HCG crude product.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104479005A (en) * | 2014-09-05 | 2015-04-01 | 上海丽珠制药有限公司 | Gonadotropin purification method |
| CN105085660A (en) * | 2015-08-21 | 2015-11-25 | 江西浩然生物医药有限公司 | Method for extracting chorionic gonadotrophin crude product from large-volume pregnant woman urine based on ultrafiltration concentration |
| CN107312081A (en) * | 2017-08-02 | 2017-11-03 | 临沂华业生物制品有限公司 | A kind of preparation method of HMG crude products |
| CN114313347A (en) * | 2022-03-02 | 2022-04-12 | 山东滨渤生物制品有限公司 | Intelligent quick collecting system of HCG crude raw materials |
| CN114736285A (en) * | 2022-05-18 | 2022-07-12 | 江苏尤里卡生物科技有限公司 | Preparation device and preparation method of human chorionic gonadotropin |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1302818A (en) * | 2000-12-12 | 2001-07-11 | 上海惠海生化制品厂 | Human chorionic gonadotropin and its preparing process |
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2013
- 2013-04-09 CN CN201310120891.XA patent/CN103193880B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1302818A (en) * | 2000-12-12 | 2001-07-11 | 上海惠海生化制品厂 | Human chorionic gonadotropin and its preparing process |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104479005A (en) * | 2014-09-05 | 2015-04-01 | 上海丽珠制药有限公司 | Gonadotropin purification method |
| CN105085660A (en) * | 2015-08-21 | 2015-11-25 | 江西浩然生物医药有限公司 | Method for extracting chorionic gonadotrophin crude product from large-volume pregnant woman urine based on ultrafiltration concentration |
| CN107312081A (en) * | 2017-08-02 | 2017-11-03 | 临沂华业生物制品有限公司 | A kind of preparation method of HMG crude products |
| CN114313347A (en) * | 2022-03-02 | 2022-04-12 | 山东滨渤生物制品有限公司 | Intelligent quick collecting system of HCG crude raw materials |
| CN114313347B (en) * | 2022-03-02 | 2022-07-15 | 山东滨渤生物制品有限公司 | Intelligent quick collecting system of HCG crude raw materials |
| CN114736285A (en) * | 2022-05-18 | 2022-07-12 | 江苏尤里卡生物科技有限公司 | Preparation device and preparation method of human chorionic gonadotropin |
| CN114736285B (en) * | 2022-05-18 | 2023-11-03 | 江苏尤里卡生物科技有限公司 | Device and method for preparing human chorionic gonadotrophin |
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| Publication number | Publication date |
|---|---|
| CN103193880B (en) | 2015-04-15 |
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